Tetrasubstituted 2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolines and method of reducing TNFα levels

ABSTRACT

Tetrasubstituted 1-oxo-2-(2,6-dioxopiperidin-3-yl)isoindolines reduce the levels of TNFα in a mammal. A typical embodiment is 1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline.

The present invention relates to tetrasubstituted2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolines, the method of reducinglevels of tumor necrosis factor α in a mammal through the administrationthereof, and pharmaceutical compositions of such tetrasubstitutedindoline derivatives.

BACKGROUND OF THE INVENTION

Tumor necrosis factor α, or TNFα, is a cytokine which is releasedprimarily by mononuclear phagocytes in response to a numberimmunostimulators. When administered to animals or humans, it causesinflammation, fever, cardiovascular effects, hemorrhage, coagulation,and acute phase responses similar to those seen during acute infectionsand shock states. Excessive or unregulated TNFα production thus has beenimplicated in a number of disease conditions. These include endotoxemiaand/or toxic shock syndrome {Tracey et al., Nature 330, 662-664 (1987)and Hinshaw et al., Circ. Shock 30, 279-292 (1990)}; cachexia {Dezube etal., Lancet, 335 (8690), 662 (1990)} and Adult Respiratory DistressSyndrome where TNFα concentration in excess of 12,000 pg/mL have beendetected in pulmonary aspirates from ARDS patients {Millar et al.,Lancet 2(8665), 712-714 (1989)}. Systemic infusion of recombinant TNFαalso resulted in changes typically seen in ARDS {Ferrai-Baliviera etal., Arch. Surg. 124(12), 1400-1405 (1989)}.

TNFα appears to be involved in bone resorption diseases, includingarthritis. When activated, leukocytes will produce bone-resorption, anactivity to which the data suggest TNFα contributes. {Bertolini et al.,Nature 319, 516-518 (1986) and Johnson et al., Endocrinology 124(3),1424-1427 (1989).} TNFα also has been shown to stimulate bone resorptionand inhibit bone formation in vitro and in vivo through stimulation ofosteoclast formation and activation combined with inhibition ofosteoblast function. Although TNFα may be involved in many boneresorption diseases, including arthritis, the most compelling link withdisease is the association between production of TNFα by tumor or hosttissues and malignancy associated hypercalcemia {Calci. Tissue Int. (US)46(Suppl.), S3-10 (1990)}. In Graft versus Host Reaction, increasedserum TNFα levels have been associated with major complication followingacute allogenic bone marrow transplants {Holler et al., Blood, 75(4),1011-1016 (1990)}.

Cerebral malaria is a lethal hyperacute neurological syndrome associatedwith high blood levels of TNFα and the most severe complicationoccurring in malaria patients. Levels of serum TNFα correlated directlywith the severity of disease and the prognosis in patients with acutemalaria attacks {Grau et al., N. Engl. J. Med. 320(24), 1586-1591(1989)}.

TNFα also plays a role in the area of chronic pulmonary inflammatorydiseases. The deposition of silica particles leads to silicosis, adisease of progressive respiratory failure caused by a fibroticreaction. Antibody to TNFα completely blocked the silica-induced lungfibrosis in mice {Pignet et al., Nature, 344:245-247 (1990)}. Highlevels of TNFα production (in the serum and in isolated macrophages)have been demonstrated in animal models of silica and asbestos inducedfibrosis {Bissonnette et al., Inflammation 13(3), 329-339 (1989)}.Alveolar macrophages from pulmonary sarcoidosis patients have also beenfound to spontaneously release massive quantities of TNFα as comparedwith macrophages from normal donors {Baughman et al., J. Lab. Clin. Med.115(1), 36-42 (1990)}.

TNFα is also implicated in the inflammatory response which followsreperfusion, called reperfusion injury, and is a major cause of tissuedamage after loss of blood flow {Vedder et al., PNAS 87, 2643-2646(1990)}. TNFα also alters the properties of endothelial cells and hasvarious pro-coagulant activities, such as producing an increase intissue factor pro-coagulant activity and suppression of theanticoagulant protein C pathway as well as down-regulating theexpression of thrombomodulin {Sherry et al., J. Cell Biol. 107,1269-1277 (1988)}. TNFα has pro-inflammatory activities which togetherwith its early production (during the initial stage of an inflammatoryevent) make it a likely mediator of tissue injury in several importantdisorders including but not limited to, myocardial infarction, strokeand circulatory shock. Of specific importance may be TNFα-inducedexpression of adhesion molecules, such as intercellular adhesionmolecule (ICAM) or endothelial leukocyte adhesion molecule (ELAM) onendothelial cells {Munro et al., Am. J. Path. 135(1), 121-132 (1989)}.

Moreover, it now is known that TNFα is a potent activator of retrovirusreplication including activation of HIV-1. {Duh et al., Proc. Nat. Acad.Sci. 86, 5974-5978 (1989); Poll et al., Proc. Nat. Acad Sci. 87, 782-785(1990); Monto et al., Blood 79, 2670 (1990); Clouse et al., J. Immunol.142, 431-438 (1989); Poll et al., AIDS Res. Hum. Retrovirus, 191-197(1992)}. AIDS results from the infection of T lymphocytes with HumanImmunodeficiency Virus (HIV). At least three types or strains of HIVhave been identified, i.e., HIV-1, HIV-2 and HIV-3. As a consequence ofHIV infection, T-cell mediated immunity is impaired and infectedindividuals manifest severe opportunistic infections and/or unusualneoplasms. HIV entry into the T lymphocyte requires T lymphocyteactivation. Other viruses, such as HIV-1, HIV-2 infect T lymphocytesafter T cell activation and such virus protein expression and/orreplication is mediated or maintained by such T cell activation. Once anactivated T lymphocyte is infected with HIV, the T lymphocyte mustcontinue to be maintained in an activated state to permit HIV geneexpression and/or HIV replication. Cytokines, specifically TNFα, areimplicated in activated T-cell mediated HIV protein expression and/orvirus replication by playing a role in maintaining T lymphocyteactivation. Therefore, interference with cytokine activity such as byprevention or inhibition of cytokine production, notably TNFα, in anHIV-infected individual assists in limiting the maintenance of Tlymphocyte caused by HIV infection.

Monocytes, macrophages, and related cells, such as kupffer and glialcells, also have been implicated in maintenance of the HIV infection.These cells, like T cells, are targets for viral replication and thelevel of viral replication is dependent upon the activation state of thecells. {Rosenberg et al., The Immunopathogenesis of HIV Infection,Advances in Immunology, 57 (1989)}. Cytokines, such as TNFα, have beenshown to activate HIV replication in monocytes and/or macrophages {Poliet al. Proc. Natl. Acad. Sci., 87, 782-784 (1990)}, therefore,prevention or inhibition of cytokine production or activity aids inlimiting HIV progression for T cells. Additional studies have identifiedTNFα as a common factor in the activation of HIV in vitro and hasprovided a clear mechanism of action via a nuclear regulatory proteinfound in the cytoplasm of cells (Osborn, et al., PNAS 86 2336-2340).This evidence suggests that a reduction of TNFα synthesis may have anantiviral effect in HIV infections, by reducing the transcription andthus virus production.

AIDS viral replication of latent HIV in T cell and macrophage lines canbe induced by TNFα {Folks et al., PNAS 86, 2365-2368 (1989)}. Amolecular mechanism for the virus inducing activity is suggested byTNFα's ability to activate a gene regulatory protein (NFκB) found in thecytoplasm of cells, which promotes HIV replication through binding to aviral regulatory gene sequence (LTR) {Osborn et al., PNAS 86, 2336-2340(1989)}. TNFα in AIDS associated cachexia is suggested by elevated serumTNFα and high levels of spontaneous TNFα production in peripheral bloodmonocytes from patients {Wright et al. J. Immunol. 141(1), 99-104(1988)}. TNFα has been implicated in various roles with other viralinfections, such as the cytomegalia virus (CMV), influenza virus,adenovirus, and the herpes family of viruses for similar reasons asthose noted.

The nuclear factor κB (NFκB) is a pleiotropic transcriptional activator(Lenardo, et al. Cell 1989, 58, 227-29). NFκB has been implicated as atranscriptional activator in a variety of disease and inflammatorystates and is thought to regulate cytokine levels including but notlimited to TNFα and also to be an activator of HIV transcription(Dbaibo, et al. J. Biol. Chem. 1993, 17762-66; Duh et al. Proc. Natl.Acad. Sci. 1989, 86, 5974-78; Bachelerie et al. Nature 1991, 350,709-12; Boswas et al. J. Acquired Immune Deficiency Syndrome 1993, 6,778-786; Suzuki et al. Biochem. And Biophys. Res. Comm. 1993, 193,277-83; Suzuki et al. Biochem. And Biophys. Res Comm. 1992, 189,1709-15; Suzuki et al. Biochem. Mol. Bio. Int. 1993, 31(4), 693-700;Shakhov et al. Proc. Natl. Acad Sci. USA 1990, 171, 35-47; and Staal etal. Proc. Natl. Acad Sci. USA 1990, 87, 9943-47). Thus, inhibition ofNFκB binding can regulate transcription of cytokine gene(s) and throughthis modulation and other mechanisms be useful in the inhibition of amultitude of disease states. The compounds described herein can inhibitthe action of NFκB in the nucleus and thus are useful in the treatmentof a variety of diseases including but not limited to rheumatoidarthritis, rheumatoid spondylitis, osteoarthritis, other arthriticconditions, septic shock, septis, endotoxic shock, graft versus hostdisease, wasting, Crohn's disease, ulcerative colitis, multiplesclerosis, systemic lupus erythrematosis, ENL in leprosy, HIV, AIDS, andopportunistic infections in AIDS. TNFα and NFκB levels are influenced bya reciprocal feedback loop. As noted above, the compounds of the presentinvention affect the levels of both TNFα and NFκB.

Many cellular functions are mediated by levels of adenosine 3',5'-cyclicmonophosphate (cAMP). Such cellular functions can contribute toinflammatory conditions and diseases including asthma, inflammation, andother conditions (Lowe and Cheng, Drugs of the Future, 17(9), 799-807,1992). It has been shown that the elevation of cAMP in inflammatoryleukocytes inhibits their activation and the subsequent release ofinflammatory mediators, including TNFα and NFκB. Increased levels ofcAMP also leads to the relaxation of airway smooth muscle.

Decreasing TNFα levels and/or increasing cAMP levels thus constitutes avaluable therapeutic strategy for the treatment of many inflammatory,infectious, immunological or malignant diseases. These include but arenot restricted to septic shock, sepsis, endotoxic shock, hemodynamicshock and sepsis syndrome, post ischemic reperfusion injury, malaria,mycobacterial infection, meningitis, psoriasis, congestive heartfailure, fibrotic disease, cachexia, graft rejection, cancer, autoimmunedisease, opportunistic infections in AIDS, rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis, other arthritic conditions,Crohn's disease, ulcerative colitis, multiple sclerosis, systemic lupuserythrematosis, ENL in leprosy, radiation damage, and hyperoxic alveolarinjury. Prior efforts directed to the suppression of the effects of TNFαhave ranged from the utilization of steroids such as dexamethasone andprednisolone to the use of both polyclonal and monoclonal antibodies{Beutler et al., Science 234, 470-474 (1985); WO 92/113 83}.

DETAILED DESCRIPTION

The present invention is based on the discovery that a class ofnon-polypeptide compounds more fully described herein decrease thelevels of TNFα and elevate the levels of adenosine 3',5'-cyclicmonophosphate.

In particular, the invention pertains to compounds of the formula:##STR1## in which each of R¹, R², R³, and R⁴, independently of theothers, is halo, alkyl of 1 to 4 carbon atoms, or alkoxy of 1 to 4carbon atoms.

The term alkyl denotes a univalent saturated branched or straighthydrocarbon chain containing from 1 to 4 carbon atoms. Representative ofsuch alkyl groups are methyl, ethyl, propyl, isopropyl, butyl. isobutyl,sec-butyl, and tert-butyl. Alkoxy refers to an alkyl group bound to theremainder of the molecule through an ethereal oxygen atom.Representative of such alkoxy groups are methoxy, ethoxy, propoxy,isopropoxy, butoxy. isobutoxy, sec-butoxy, and tert-butoxy. PreferablyR¹, R², R³, and R⁴ are chloro, fluoro, methyl or methoxy.

The compounds of Formula I are used, under the supervision of qualifiedprofessionals, to inhibit the undesirable effects of TNFα. The compoundscan be administered orally, rectally, or parenterally, alone or incombination with other therapeutic agents including antibiotics,steroids, etc., to a mammal in need of treatment.

The compounds of the present invention also can be used topically in thetreatment or prophylaxis of topical disease states mediated orexacerbated by excessive TNFα production, respectively, such as viralinfections, such as those caused by the herpes viruses, or viralconjunctivitis, etc.

The compounds also can be used in the veterinary treatment of mammalsother than humans in need of prevention or inhibition of TNFαproduction. TNFα mediated diseases for treatment, therapeutically orprophylactically, in animals include disease states such as those notedabove, but in particular viral infections. Examples include felineimmunodeficiency virus, equine infectious anaemia virus, caprinearthritis virus, visna virus, and maedi virus, as well as otherlentiviruses.

The compounds can be prepared using methods which are known in general.In particular, the compounds can be prepared through the reaction of2,6-dioxopiperidin-3-ammonium chloride, and a lower alkyl ester of2-bromomethyl-3,4,5,6-tetrasubstituted-benzoic acid in the presence ofan acid acceptor such as dimethylaminopyridine or triethylamine.##STR2##

The tetrasubstituted benzoate intermediates are known or can be obtainedthough conventional processes. For example, a lower alkyl ester of a3,4,5,6-tetrasubstituted ortho-toluic acid is brominated withN-bromosuccinimide under the influence of light to yield the lower alkyl2-(bromomethyl)-3,4,5,6-tetrasubstitutedbenzoate.

Alternatively, a dialdehyde is allowed to react with2,6-dioxopiperidin-3-ammonium chloride: ##STR3##

In a further method, an appropriatedly substituted phthalidimideintermediate is selectively reduced: ##STR4##

Finally, a dialdehyde is allowed to react with glutamine and theresulting 2-(1-oxo-isoindolin-2-yl)glutaric acid then cyclized to yielda 4,5,6,7-tetrasubstituted 1-oxo-2-(2,6-dioxopiperidin-3-yl)-isoindoline of Formula I: ##STR5##

The compounds of Formula I possess a center of chirality and can existas optical isomers. Both the racemates of these isomers and theindividual isomers themselves, as well as diastereomers when there aretwo chiral centers, are within the scope of the present invention. Theracemates can be used as such or can be separated into their individualisomers mechanically as by chromatography using a chiral absorbant.Alternatively, the individual isomers can be prepared in chiral form orseparated chemically from a mixture by forming salts with a chiral acid,such as the individual enantiomers of 10-camphorsulfonic acid, camphoricacid, α-bromocamphoric acid, methoxyacetic acid, tartaric acid,diacetyltartaric acid, malic acid, pyrrolidone-5-carboxylic acid, andthe like, and then freeing one or both of the resolved bases, optionallyrepeating the process, so as obtain either or both substantially free ofthe other; i.e., in a form having an optical purity of >95%.

The present invention also pertains to the physiologically acceptablenon-toxic acid addition salts of the compounds of Formula I. Such saltsinclude those derived from organic and inorganic acids such as, withoutlimitation, hydrochloric acid, hydrobromic acid, phosphoric acid,sulfuric acid, methanesulphonic acid, acetic acid, tartaric acid, lacticacid, succinic acid, citric acid, malic acid, maleic acid, sorbic acid,aconitic acid, salicylic acid, phthalic acid, embonic acid, enanthicacid, and the like.

Oral dosage forms include tablets, capsules, dragees, and similarshaped, compressed pharmaceutical forms containing from 1 to 100 mg ofdrug per unit dosage. Isotonic saline solutions containing from 20 to100 mg/mL can be used for parenteral administration which includesintramuscular, intrathecal, intravenous and intra-arterial routes ofadministration. Rectal administration can be effected through the use ofsuppositories formulated from conventional carriers such as cocoabutter.

Pharmaceutical compositions thus comprise one or more compounds ofFormula I associated with at least one pharmaceutically acceptablecarrier, diluent or excipient. In preparing such compositions, theactive ingredients are usually mixed with or diluted by an excipient orenclosed within such a carrier which can be in the form of a capsule orsachet. When the excipient serves as a diluent, it may be a solid,semi-solid, or liquid material which acts as a vehicle, carrier, ormedium for the active ingredient. Thus, the compositions can be in theform of tablets, pills, powders, elixirs, suspensions, emulsions,solutions, syrups, soft and hard gelatin capsules, suppositories,sterile injectable solutions and sterile packaged powders. Examples ofsuitable excipients include lactose, dextrose, sucrose, sorbitol,mannitol, starch, gum acacia, calcium silicate, microcrystallinecellulose, polyvinylpyrrolidinone, cellulose, water, syrup, and methylcellulose, the formulations can additionally include lubricating agentssuch as talc, magnesium stearate and mineral oil, wetting agents,emulsifying and suspending agents, preserving agents such as methyl- andpropylhydroxybenzoates, sweetening agents or flavoring agents.

The compositions preferably are formulated in unit dosage form, meaningphysically discrete units suitable as a unitary dosage, or apredetermined fraction of a unitary dose to be administered in a singleor multiple dosage regimen to human subjects and other mammals, eachunit containing a predetermined quantity of active material calculatedto produce the desired therapeutic effect in association with a suitablepharmaceutical excipient. The compositions can be formulated so as toprovide an immediate, sustained or delayed release of active ingredientafter administration to the patient by employing procedures well knownin the art.

Oral dosage forms include tablets, capsules, dragees, and similarshaped, compressed pharmaceutical forms containing from 1 to 100 mg ofdrug per unit dosage. Isotonic saline solutions containing from 20 to100 mg/mL can be used for parenteral administration which includesintramuscular, intrathecal, intravenous and intra-arterial routes ofadministration. Rectal administration can be effected through the use ofsuppositories formulated from conventional carriers such as cocoabutter.

Pharmaceutical compositions thus comprise one or more compounds ofFormula I associated with at least one pharmaceutically acceptablecarrier, diluent or excipient. In preparing such compositions, theactive ingredients are usually mixed with or diluted by an excipient orenclosed within such a carrier which can be in the form of a capsule orsachet. When the excipient serves as a diluent, it may be a solid,semi-solid, or liquid material which acts as a vehicle, carrier, ormedium for the active ingredient. Thus, the compositions can be in theform of tablets, pills, powders, elixirs, suspensions, emulsions,solutions, syrups, soft and hard gelatin capsules, suppositories,sterile injectable solutions and sterile packaged powders. Examples ofsuitable excipients include lactose, dextrose, sucrose, sorbitol,mannitol, starch, gum acacia, calcium silicate, microcrystallinecellulose, polyvinylpyrrolidinone, cellulose, water, syrup, and methylcellulose, the formulations can additionally include lubricating agentssuch as talc, magnesium stearate and mineral oil, wetting agents,emulsifying and suspending agents, preserving agents such as methyl- andpropylhydroxybenzoates, sweetening agents or flavoring agents.

The compositions preferably are formulated in unit dosage form, meaningphysically discrete units suitable as a unitary dosage, or apredetermined fraction of a unitary dose to be administered in a singleor multiple dosage regimen to human subjects and other mammals, eachunit containing a predetermined quantity of active material calculatedto produce the desired therapeutic effect in association with a suitablepharmaceutical excipient. The compositions can be formulated so as toprovide an immediate, sustained or delayed release of active ingredientafter administration to the patient by employing procedures well knownin the art.

The following examples will serve to further typify the nature of thisinvention but should not be construed as a limitation in the scopethereof, which scope is defined solely by the appended claims.

EXAMPLE 1

A mixture of 16.25 g of 2,6-dioxopiperidin-3-ammonium chloride, and 30.1g of methyl 2-bromomethyl-3,4,5,6-tetrafluorobenzoate, and 12.5 g oftriethylamine in 100 mL of dimethylformamide is stirred at roomtemperature for 15 hours. The mixture is then concentrated in vacuo andthe residue mixed with methylene chloride and water. The aqueous layeris separated and back-extracted with methylene chloride. The combinedmethylene chloride solutions are dried over magnesium sulfate andconcentrated in vacuo to give1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline.

In a similar fashion1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrachloroisoindoline,1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethylisoindoline, and1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethoxyisoindoline areobtained by substituting equivalent amounts of2-bromomethyl-3,4,5,6-tetrachlorobenzoate,2-bromomethyl-3,4,5,6-tetramethylbenzoate, and2-bromomethyl-3,4,5,6-tetramethoxybenzoate, respectively, for2-bromomethyl-3,4,5,6-tetrafluorobenzoate.

EXAMPLE 2

Tablets, each containing 50 mg of1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline, can beprepared in the following manner:

    ______________________________________                                        Constituents (for 1000 tablets)                                               ______________________________________                                        1-oxo-2-(2,6-dioxo-                                                                             50.0 g                                                      piperidin-3-yl)-4,5,6,7-                                                      tetrafluoroisoindoline                                                        lactose           50.7 g                                                      wheat starch      7.5 g                                                       polyethylene glycol 6000                                                                        5.0 g                                                       talc              5.0 g                                                       magnesium stearate                                                                              1.8 g                                                       demineralized water                                                                             q.s.                                                        ______________________________________                                    

The solid ingredients are first forced through a sieve of 0.6 mm meshwidth. The active ingredient, lactose, talc, magnesium stearate and halfof the starch then are mixed. The other half of the starch is suspendedin 40 mL of water and this suspension is added to a boiling solution ofthe polyethylene glycol in 100 mL of water. The resulting paste is addedto the pulverulent substances and the mixture is granulated, ifnecessary with the addition of water. The granulate is dried overnightat 35° C., forced through a sieve of 1.2 mm mesh width and compressed toform tablets of approximately 6 mm diameter which are concave on bothsides.

EXAMPLE 3

Tablets, each containing 100 mg of1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrachloroisoindoline, can beprepared in the following manner:

    ______________________________________                                        Constituents (for 1000 tablets)                                               ______________________________________                                        1-oxo-2-(2,6-dioxopiperidin-3-yl)-                                                                 100.0 g                                                  4,5,6,7-tetrachloroisoindoline                                                lactose              100.0 g                                                  wheat starch          47.0 g                                                  magnesium stearate    3.0 g                                                   ______________________________________                                    

All the solid ingredients are first forced through a sieve of 0.6 mmmesh width. The active ingredient, lactose, magnesium stearate and halfof the starch then are mixed. The other half of the starch is suspendedin 40 mL of water and this suspension is added to 100 mL of boilingwater. The resulting paste is added to the pulverulent substances andthe mixture is granulated, if necessary with the addition of water. Thegranulate is dried overnight at 35° C., forced through a sieve of 1.2 mmmesh width and compressed to form tablets of approximately 6 mm diameterwhich are concave on both sides.

EXAMPLE 4

Tablets for chewing, each containing 75 mg of1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline, can beprepared in the following manner:

    ______________________________________                                        Composition (for 1000 tablets)                                                ______________________________________                                        1-oxo-2-(2,6-dioxo- 75.0 g                                                    piperidin-3-yl)-4,5,6,7-tetra-                                                fluoroisoindoline                                                             mannitol            230.0 g                                                   lactose             150.0 g                                                   talc                21.0 g                                                    glycine             12.5 g                                                    stearic acid        10.0 g                                                    saccharin            1.5 g                                                    5% gelatin solution q.s.                                                      ______________________________________                                    

All the solid ingredients are first forced through a sieve of 0.25 mmmesh width. The mannitol and the lactose are mixed, granulated with theaddition of gelatin solution, forced through a sieve of 2 mm mesh width,dried at 50° C. and again forced through a sieve of 1.7 mm mesh width.1-Oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline, theglycine and the saccharin are carefully mixed, the mannitol, the lactosegranulate, the stearic acid and the talc are added and the whole ismixed thoroughly and compressed to form tablets of approximately 10 mmdiameter which are concave on both sides and have a breaking groove onthe upper side.

EXAMPLE 5

Tablets, each containing 10 mg of1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethylisoindoline, can beprepared in the following manner:

    ______________________________________                                        Composition (for 1000 tablets)                                                ______________________________________                                        1-oxo-2-(2,6-dioxo-                                                                             10.0 g                                                      piperidin-3-yl)-4,5,6,7-                                                      tetramethylisoindoline                                                        lactose           328.5 g                                                     corn starch       17.5 g                                                      polyethylene glycol 6000                                                                         5.0 g                                                      talc              25.0 g                                                      magnesium stearate                                                                               4.0 g                                                      demineralized water                                                                             q.s.                                                        ______________________________________                                    

The solid ingredients are first forced through a sieve of 0.6 mm meshwidth. Then the active imide ingredient, lactose, talc, magnesiumstearate and half of the starch are intimately mixed. The other half ofthe starch is suspended in 65 mL of water and this suspension is addedto a boiling solution of the polyethylene glycol in 260 mL of water. Theresulting paste is added to the pulverulent substances, and the whole ismixed and granulated, if necessary with the addition of water. Thegranulate is dried overnight at 35° C., forced through a sieve of 1.2 mmmesh width and compressed to form tablets of approximately 10 mmdiameter which are concave on both sides and have a breaking notch onthe upper side.

EXAMPLE 6

Gelatin dry-filled capsules, each containing 100 mg of1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethoxyisoindoline, canbe prepared in the following manner:

    ______________________________________                                        Composition (for 1000 capsules)                                               ______________________________________                                        1-oxo-2-(2,6-dioxopiperidin-3-                                                                    100.0 g                                                   yl)-4,5,6,7-tetramethoxy-                                                     isoindoline                                                                   microcrystalline cellulose                                                                        30.0 g                                                    sodium lauryl sulfate                                                                             2.0 g                                                     magnesium stearate  8.0 g                                                     ______________________________________                                    

The sodium lauryl sulfate is sieved into the1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethoxyisoindolinethrough a sieve of 0.2 mm mesh width and the two components areintimately mixed for 10 minutes. The microcrystalline cellulose is thenadded through a sieve of 0.9 mm mesh width and the whole is againintimately mixed for 10 minutes. Finally, the magnesium stearate isadded through a sieve of 0.8 mm width and, after mixing for a further 3minutes, the mixture is introduced in portions of 140 mg each into size0 (elongated) gelatin dry-fill capsules.

EXAMPLE 7

A 0.2% injection or infusion solution can be prepared, for example, inthe following manner:

    ______________________________________                                        1-oxo-2-(2,6-dioxopiperidin-3-yl)-                                                                   5.0      g                                             4,5,6,7-tetrafluoroisoindoline                                                sodium chloride        22.5     g                                             phosphate buffer pH 7.4                                                                              300.0    g                                             demineralized water    to 2500.0                                                                              mL                                            ______________________________________                                    

1-Oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline isdissolved in 1000 mL of water and filtered through a microfilter. Thebuffer solution is added and the whole is made up to 2500 mL with water.To prepare dosage unit forms, portions of 1.0 or 2.5 mL each areintroduced into glass ampoules (each containing respectively 2.0 or 5.0mg of imide).

What is claimed is:
 1. A compound of the formula: ##STR6## in which eachof R¹, R², R³, and R⁴, independently of the others, is halo, alkyl of 1to 4 carbon atoms, or alkoxy of 1 to 4 carbon atoms.
 2. The compoundaccording to claim 1 in which R¹, R², R³, and R⁴ are the same and eachis chloro, fluoro, methyl, or methoxy.
 3. The compound according toclaim 2 which is1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrafluoroisoindoline.
 4. Thecompound according to claim 2 which is1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetrachloroisoindoline.
 5. Thecompound according to claim 2 which is1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethylisoindoline.
 6. Thecompound according to claim 2 which is1-oxo-2-(2,6-dioxopiperidin-3-yl)-4,5,6,7-tetramethoxyisoindoline. 7.The method of reducing undesirable levels of TNFα in a mammal whichcomprises administering thereto an effective amount of a compoundaccording to claim
 1. 8. A pharmaceutical composition comprising aquantity of a compound according to claim 1 sufficient uponadministration in a single or multiple dose regimen to reduce levels ofTNFα in a mammal in combination with a carrier.